Pest resistant plants

Abstract

An isolated nucleic acid molecule encoding a 7-epizingiberene synthase, a chimeric gene comprising the nucleic acid molecule, vectors comprising the same, as well as isolated 7-epizingiberene synthase proteins themselves, are provided. In addition, transgenic plants and plant cells comprising a gene encoding a 7-epizingiberene synthase, optionally integrated in its genome, and methods for making such plants and cells, are provided. Especially Solanaceae plants and plant parts (seeds, fruit, leaves, etc.) with enhanced insect pest resistance are provided.

The invention claimed is:  
     
       1. A method for preparing 7-epizingiberene and/or R-curcumene comprising:
 a) transforming a host cell with a recombinant nucleic acid molecule comprising a nucleic acid sequence encoding a polypeptide comprising the amino acid sequence of SEQ ID NO: 1 or an amino acid sequence at least 99% identical to the full length of SEQ ID NO: 1, wherein in the transformed host cell the nucleic acid sequence encoding the polypeptide is operably linked to an active promoter; and 
 b) culturing said host cell under conditions permitting production of 7-epizingiberene and/or R-curcumene. 
 
     
     
       2. A method for producing 7-epizingiberene from zFPP in a host cell, comprising:
 a) introducing into said host cell a first nucleic acid sequence encoding a polypeptide comprising the amino acid sequence of SEQ ID NO:6 or an amino acid sequence at least 95% identical to the full length of SEQ ID NO:6 over the entire length, and a second nucleic acid sequence encoding a polypeptide comprising the amino acid sequence of SEQ ID NO: 1 or an amino acid sequence at least 99% identical to the full length of SEQ ID NO:1, wherein in the host cell obtained then first nucleic acid sequence and second nucleic acid sequence are operably linked to an active promoter; 
 b) culturing the host cell in suitable conditions for the expression of said first and said second nucleic acid sequences; and, 
 c) collecting the 7-epizingiberene contained in said cell and/or in a culture medium thereof. 
 
     
     
       3. A method for producing a transgenic plant having enhanced insect pest resistance compared to a non-transgenic control plant, said method comprising the steps of:
 (a) transforming a plant or plant cell with a nucleic acid molecule encoding a polypeptide comprising the amino acid sequence of SEQ ID NO: 1 or an amino acid sequence at least 99% identical to the full length of SEQ ID NO:1, operably linked to a promoter active in plant cells, and 
 (b) regenerating a plant. 
 
     
     
       4. The method according to  claim 3 , wherein said nucleic acid molecule is integrated into the genome of said plant. 
     
     
       5. The method according to  claim 3 , further comprising the step of
 (c) creening the regenerated plant, or a plant derived therefrom by selfing or crossing, for resistance to one or more insect pests and identifying a plant comprising enhanced resistance to one or more of said insect pests. 
 
     
     
       6. The method according to  claim 3 , wherein said promoter is an insect pest inducible promoter. 
     
     
       7. The method according to  claim 3 , wherein the plant belongs to the family Solanaceae. 
     
     
       8. The method according to  claim 7 , wherein the plant is of the genus  Solanum.    
     
     
       9. The method according to  claim 3 , wherein the method further comprises the step of:
 transforming the plant or plant cell with a nucleic acid molecule encoding the amino acid sequence of SEQ ID NO:6 or an amino acid sequence comprising at least 80% amino acid sequence identity to the amino acid sequence of SEQ ID NO:6, over the entire length, operably linked to a promoter active in plant cells. 
 
     
     
       10. The method according to  claim 1 , further comprising the step of c) isolating the 7-epizingiberene produced in step b). 
     
     
       11. The method according to  claim 10 , further comprising the steps of d) dehydrogenating the 7-epizingiberene to produce R-curcumene. 
     
     
       12. The method according to  claim 1 , wherein the nucleic acid sequence is operably linked to a heterologous promoter in the transformed host cell. 
     
     
       13. The method according to  claim 1 , wherein the nucleic acid sequence encodes a polypeptide comprising the amino acid sequence of SEQ ID NO: 1. 
     
     
       14. The method according to  claim 1 , wherein the recombinant nucleic acid molecule comprises a cDNA sequence that encodes a polypeptide comprising the amino acid sequence of SEQ ID NO: 1 or an amino acid sequence at least 99% identical to the full length of SEQ ID NO:1. 
     
     
       15. The method according to  claim 2 , wherein the second nucleic acid sequence is operably linked to a heterologous promoter in the transformed host cell. 
     
     
       16. The method according to  claim 2 , wherein the first nucleic acid sequence encodes a polypeptide comprising the amino acid sequence of SEQ ID NO: 6, and the second nucleic acid sequence encodes a polypeptide comprising the amino acid sequence of SEQ ID NO: 1. 
     
     
       17. The method according to  claim 2 , wherein the second nucleic acid sequence comprises a cDNA sequence that encodes a polypeptide comprising the amino acid sequence of SEQ ID NO: 1 or an amino acid sequence at least 99% identical to the full length of SEQ ID NO:1. 
     
     
       18. A method for preparing 7-epizingiberene and/or R-curcumene comprising the steps of: a) culturing a host cell transformed with a recombinant nucleic acid molecule comprising a nucleic acid sequence that encodes a polypeptide comprising the amino acid sequence of SEQ ID NO: 1 or an amino acid sequence at least 99% identical to the full length of SEQ ID NO: 1, wherein in the host cell the nucleic acid sequence encoding the polypeptide is operably linked to an active promoter, under conditions permitting production of 7-epizingiberene and/or R-curcumene. 
     
     
       19. The method according to  claim 18 , further comprising the step of b) isolating the 7-epizingiberene produced in step a). 
     
     
       20. The method according to  claim 19 , further comprising the steps of c) dehydrogenating the 7-epizingiberene to produce R-curcumene. 
     
     
       21. A method for preparing 7-epizingiberene and/or R-curcumene comprising the steps of:
 a) heterologously expressing in a host cell a polypeptide comprising the amino acid sequence of SEQ ID NO: 1 or an amino acid sequence at least 99% identical to the full length of SEQ ID NO: 1, under conditions permitting production of 7-epizingiberene. 
 
     
     
       22. The method according to  claim 21 , further comprising the step of b) isolating the 7-epizingiberene produced in step a). 
     
     
       23. The method according to  claim 22 , further comprising the steps of c) dehydrogenating the 7-epizingiberene to produce R-curcumene.